The linkage between Qi and mitochondria was investigated by exploring the effect of Traditional Chinese Medicine (TCM) Qi-invigorating herbs on mitochondrial function at the biochemical and molecular levels. Three Chinese herbs (<i></span><i><span style="font-family:Verdana;">Astragali radix</span></i><span style="font-family:Verdana;">,</span><i><span style="font-family:Verdana;"> Herba cistanche</span></i><span style="font-family:Verdana;">, </span><i><span style="font-family:Verdana;">Panax ginseng</span></i><span style="font-family:Verdana;"></i>) were used to treat cultured mouse kidney cells and the generation of adenosine triphosphate (ATP) was measured. The Qi-invigorating herb, <i></span><i><span style="font-family:Verdana;">Astragali radix</span></i><span style="font-family:Verdana;"></i>, was selected for further </span></span><span style="font-family:""><span style="font-family:Verdana;">study using additional biological and molecular parameters, including ATP, reactive oxygen species (ROS), mitochondrial membrane potential (MMP),</span><span><span style="font-family:Verdana;"> mtDNA copies, superoxide dismutase (SOD), glutathione (GSH), cell growth, cell viability and transcriptomes. We also chose two concentrations of <i></span><i><span style="font-family:Verdana;">Astragali radix</span></i><span style="font-family:Verdana;"></i> to study the hormetic effect. The results indicated that: 1) Qi-invigorating herbs have significant effects on the function of mitochondria, with ATP production and the antioxidant capacity being significantly enhanced, and ROS le</span></span><span style="font-family:Verdana;">vels being reduced, allowing for a more optimal oxidation environment. The effect of the herbs followed a hormetic curve with a stimulating effect at lower</span><span> </span><span style="font-family:Verdana;">doses but an inhibiting effect at high doses;2) The growth of the cells was not</span><span> </span><span style="font-family:Verdana;">affected despite numerous biochemical changes associated